Method of inducing simultaneous mounting of silkworms

ABSTRACT

SIMULTANEOUS MOUNTING A SILKWORMS AND CORRESPONDING IMPROVEMENT OF QUALITY OF COCOONS ARE ACHIEVED BY FEEDING THE SILKWORMS A DIET CONTAINING ABOUT 5 TO 20 GAMMAS PER GRAM OF COMPOUND(S) OF THE FORMULA   2,3,14-TRI(HO-),17-R-ANDROST-7-EN-6-ONE   WHEREIN THE 2- AND 3-POSITIONED OH GROUPS ARE A- OR BORINETED, AND R IS 1,5-DIMETHYLHEXYL SUBSTITUTED BY 2 TO 4 OH GROUPS, OR ALKANOATE(S) THEREOF WITH AT MOST 4 CARBON ATOMS, SEVERALLY OR IN ADMIXTURE.

United States Patent 3 709 990 METHOD OF lNmicnwo SIMULTANEOUS MOUNTING or SILKWORMS Masaki Kamada and Tetsuo Okauchi, Osaka, Japan,

irssignors to Takeda Chemical Industries, Ltd., Osaka,

apan

No Drawing. Continuation-impart of application Ser. No. 688,001, Dec. 5, 1967. This application May 19, 1970,

Ser. No. 38,897

Int. Cl. A61k 17/00 US. Cl. 424-243 16 Claims ABSTRACT OF THE DISCLOSURE wherein the 2- and 3-positioncd OH groups are ozor 3- oriented, and R is 1,5-dimethylhexyl substituted by 2 to 4 OH groups, or alkanoate(s) thereof with at most 4 carbon atoms, severally or in admixture.

This is a continuation-in-part application of applicants copending application Ser. No. 688,001, filed Dec. 5, 1967 and now abandoned.

This invention relates to a method for inducing simultaneous mounting of silkworms, and also relates to artificial feedstuff therefor.

When silkworrns are reared by hitherto known feedstutfs, their starting of mounting does not always Occur simultaneously and, as a result, the produced cocoons are hardly of homogeneous quality.

Further, the time lag in mounting has been a stumbling block to economy of man-power and to realization of mechanized sericulture.

The object of this invention is, therefore, a method for inducing simultaneous mounting. Another object of this invention is to provide a new artificial feedstuif by which simultaneous mounting of silkworms is induced.

The principal object is realized by feeding silkworms of the fifth instar on a compound of the formula:

3,709,990 Patented Jan. 9, 1973 ice (wherein the two hydroxyls at positions 2 and 3 are uor fi-oriented, R is 1,5-dimethylhexyl having 2 to 4 hydroxyl groups) or on a mixture of such compounds (I), in admixture with the silkworm feedstuif.

The compounds of the Formula I may be added to the feedstuif in the form of alkanoates thereof, having at most 4 carbon atoms (e.g. formates, acetates, etc.), or in a crude form containing the said compounds (e.g. extract, dried raw materials as detailed hereinafter), and can also be added either in the form of a solution or in the solid form.

These compounds have been known as moulting hormones, and their administration has been effected by injection, but there has been no information in regard to their effect on simultaneous mounting of silkworms and the fact that such effect can be brought about through oral administration.

As advantageously employed and easily available compounds of the Formula I, there are exemplified:

25,3 f1,14a,20,22-pentahydroxycholest-7-en-6-one; 2u,3a,14u,20,22-pentahydroxycholest-7-en-6-one; 23,35,l404,20,22.,24-hexahydroxycholest-7-en-6-0ne;

2 3, 318, 14a,22,25 -pentahydroxycholest-7-en-6-one; 2 3,313,14u,20,22,25-hexahydroxycholest-7-en-6-one; 2,8,35,14a,20,22,26-hexahydroxycholest-7-en-6-one;

2 5,3 13, :,20, 22,25 ,26-heptahydroxychole st-7-en-6-one.

These compounds are obtained, for example, from such plants as Podocarpus Nakaii, Podocarpus macrophyllus, Podocarpus chinensis, Achyranthes japonica Nakaii, Achyranthes radix, Achyranthes longifolia Makino, Taxus cuspidata, Afuga documbens, Ajuga nipponensis, Ajuga incisa, 'Ajwga japonica Miq., Ajuga shikotanensis Miyabe et Tatewaki, A juga yesoensz's Maxim, Stachyurus praecox, Helleborus niger, Polypodium japonicum, Polypodium niponium Mett, Struthiopterzs nipo nica, Dryopteris T helypteris, Lychnis Miqueliana, Dryopteris Thelypteris A. Gray, Lychnis Miqueliana Rohrb., Cyprus rotundus L, Siler divaricatum Benth et Hook fil, Siler Libanoz'tus, Gardenia jasminoides Ellis f. grandiflora Makino, Alisma Plantago-aquatica L. var. orientile Samuels, Glycyrrhiza glabra L, Gledtschia japonica Miq., Digenea simplex C. Agardh., Inula jwponica Thunb., Vitex magapotamica Moldenke.

The nomenclature of the above-enumerated plants is in accordance with the new plant picture book edited by Makino (published by Hokuryukan in 1951).

These raw materials contain the aforementioned compounds solely or as a mixture thereof in a concentration of about 0.005 to 0.5% by weight on the dry basis.

The compounds of the Formula I are easily isolated from the raw materials. For example, 2/3,3fl,l4oc,20,22- pentahydroxycholest-7-en-6-0ne is extracted from P0d0- carpus Nakaii leaves with alcohol. After distilling 01f the solvent, the substance is extracted again from the resultant solution with chloroform, folowed by chromatography on silica gel. The objective compound is recovered as crystals from methanol solution of the obtained eluate. M.P. 259- 260 C.; [u] =+9l (0.101% MeOH).

In analogous manner, the other aforementioned compounds I are obtained from raw materials.

The aforesaid acylates are easily synthesized by subjecting the corresponding starting materials to acylation in per se conventional manner; cf. U.S. Pat. No. 3,271,250.

There may be employed various kinds of components for feedstuif, including the said compounds and other additives.

One of the compounds I or a mixture thereof is added to feedstutf in such amount that the concentration of the said compounds is from about to about 20 gammas per gram of the feedstuff on the dry basis.

As the concentration of the compounds I in the feedstuff exceeds 20 gammas, the obtained cocoons become less in quantity and inferior in quality.

As the composition of feedstuff, there may be exemplified: diluting and nourishing materials such as agar, carboxyrnethyl cellulose, methyl cellulose, alginic acid, soy bean meal, soy bean flour, defatted soy bean meal, soy bean-curd refuse, soy bean curd, fermented soy beans, dried fermented soy bean meal, defatted dry bean paste, rice flour, wheat meal, processed grain flour, sugars (e.g. sucrose, glucose, lactose, rafiinose, fructose), straw ash, yeast or its processed matter, fish meal, etc. These also serve as carrier. Further, there may be exemplified the following nutritious components for the growth of silkworms: amino acids such as glutamic acid, aspartic acid, leucine, lysine, tyrosine, tryptophan, valine, threonine, serine, proline, glycine, alanine, isoleucine, phenylalanine, arginine, methionine and salts thereof; animal or plant proteins; vitamins such as pantothenic acid, nicotinic acid, vitamin B vitamin B vitamin B vitamin B vitamin C, vitamin D, biotin, folic acid, vitamin K, vitamin E, vitamin P, inositol, orotic acid, lipoic acid; nucleic acid and its related compounds such as adenylic acid, guanylic acid, uridylic acid, cytidylic acid, inosinic acid, desoxyadenylic acid, thymidylic acid, adenine, guanine, cytosine, hypoxanthine; inorganic salts such as sodium phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, magnesium phosphate, ammonium dihydrogen phosphate, sodium carbonate, sodium hydrogen carbonate, potassium carbonate, sodium chloride, potassium chloride, calcium chloride, polyphos- EXAMPLE 1 Following artificial redsttia'i employed; 7 V H Basal components: Part(s) by weight Water: 1.6 times as much by volume as all the other components. I

1 Composition: NaCl 0.105 KCl 0.120 KH2PO4 0.310 C8.a(PO4)2 0.149 Ca'COs 0.210 MgSCM 0.090 FBPO4A'H2O f 0.0147 M11504 0.0002 K2AIs(SO4)4 .24H2O 0.00009 Cu 4. 2O 0.00039 N aF- 0.00057 A mixture of biotin 0.2 mg., calcium pantoate 2.0 mg., folic acid 0.2 mg., nicotinic amide 2.0 mg., vitamin B0 1.0 mg., vitamin B2 1.0 mg., Vitamin B1 1.0 mg., vitamin C 39.16 mg., and carnitine 1.0 mg diluted to 1 gram with potato starch.

2fl,3,B,14m,20,22 L pentahydroxycholest 7-en-6-one is added to the basal components. Thus-obtained feedstutf is fed to silkworms (race: Gunko x Banri) from the fourth day of the final instar. The time of mounting and spinning after the start of feeding is observed. The results are shown in Table 1. Hours in the tablesin the following examples are those required for the silkworms to start mounting or spinning after the first feeding.

phoryl salt; honey or its processed matter; enzymes such as diastase, papain, trypsin; antiseptic substances such as formaldehyde, sodium dehydrogenacetate, vitamin K sodium sorbate; antibiotics and sulfa drugs such as tetracycline, chlortetracycline, oxytetracycline, 'neomycin, dextromycin, kanamycin, triochomycin, mikamycin, penicillin, sulfisomidine, sulfisoxazole, metoxypyridazine, particularly streptomycin, dihydrostreptomycin, streptothricin, chloramphenicol, oleandomycin, leucomycin, erythromycin, picromycin; chemicals such as nitrofuryl acrylic amide, hinokitiol, furazolidone, nitrofurasone, butyl phydroxybenzoate, propyl-hydroxybenzoate; others such as glucuronic acid or salt thereof v(ethyl glucuronate, etc.), glucuronic acid amide, 2 amino-Z-desoxy-D-glucuronic acid or salt thereof; N-glucuronosideglucosamine or salt thereof; chlorogenic acid, resorcylic acid, gentisic acid, homogentisic acid, piperic acid, gallic acid, cafieic acid, or salts or esters thereof; quinic acid, gulonic acid, shikimic acid, or salts or esters thereof; polyphenols (e.g. vanillin, protocatechualdehyde), etc.

The following examples are merely for illustrative purposes, and are not intended to be construed as limitations of the present invention.

TABLE 1 Amount Mounting number atter- Spinuintg numer er addition, Number 1100 g. of ofsllk- 1s a 24 42 4s 2 4s feedstuff worms hours hours hours hours hours hours 0 40 2 9 22 29 3 6 4. 8 40 B 4 11 40 1 4 24 Y 40 o s 40 4o a 29 120 40 8 36 40 40 30 40 600 40 22 4O 40 40 40 4.0 3, 000 40 40 40 40 &0 40 40 EXAMPLE 2 2,6,3/8,14a,22,25-pentahydroxycholest 7 en-6-one (referred to as compound A, M.P. 235 to 237 C. (x +58.6) or 2 6,3 3,14a,20,22,25-hexahydroxycholest-7-enone (referred to as compound B, M.P. 237.5 to 239.5 C, [a] =-{63 (0:1.0 MeOH) was added to the basal components in Example 1.

Thus obtained artificial feedstuif is fed to silkworms (race: Gunko x Banri) from the fourth day of the final instar.

The time of mounting and spinning is observed and the results on Compounds A and B are shown in Table 2 and Table 3 respectively.

TABLE 2 Amount Spinning addition. Mounting number number 71100 gi Number; at after- 0 0 feedsilk- 24 42 48 42 48 Run stuti worms hours hours hours hours hours 0 40 0 5 22 1 6 3. 2 40 0 8 18 3 ll 16 40 0 8 40 4 26 40 6 38 i0 29 38 300 40 20 40 40 36 4.0 2,000 40 40 40 40 40 40 TABLE 8 Amount Spinning addition, Mounting number number 7/100 g Number aiter after feedsilk- 24 42 48 42 48 Run stufi worms hours hours hours hours hours Control. 40 14 31 5 16 3. 2 40 4 13 40 4 14 16. 0 40 3 24 40 12 40 80 40 28 40 40 30 40 400 40 40 40 40 40 40 2, 000 40 4O 40 40 40 40 EXAMPLE 3 Dried root meal of Achyranthes japonica Nakaii is added to the basal components in Example 1.

Thus obtained artificial feedstufi is fed to silk Worms (race: Gunko x Banri) from the fifth night of the final instar. The-time of mounting and spinning is observed and the results are shown in Table 4.

Water: 1.6 times as much by volume as all the other components.

TABLE 4 Amount Mounting number after Spinning number alteraddition, Number /100 g. of of silk- 18 24 42 48 42 Run feedstufi worms hours hours hours hours hours hours Control 0 40 0 25 24 25 156 40 27 32 40 40 33 40 303 40 30 40 40 40 40 40 625 40 28 40 40 4O 40 40 1, 250 40 40 40 40 40 40 40 2, 500 40 40 40 40 40 40 40 The dried root meal contains about 1000 p.p.m. of the compounds (I) on the dry basis.

EXAMPLE 4 1000 of 2B,3,8,14oz,20,22,24 hexahydroxycholest-7- 4 en-6-0ne tetra acetate (M.P. 1l3l15 C.) is added to 100 grams of the basal components in Example 1.

Thus obtained artificial feedstuff is fed to silkworms (race: Gunko x Banri).

The silkworms are induced to mount in 16 hrs. after 2000 each of testing materials is added to 100 g. of the basal feedstuff, respectively. Thus obtained feedstufi is given to silkworms (race: Gunko x Banri) from the 6th day of the final instar for 12 hours. 7

feeding, and the time course of spinning is observed to give the results shown in Table 5.

TABLE 5 Number of spinning Worms after feeding i0r- Number Of silk- 16 20 24 28 32 38 40 44 Run worms hours hours hours hours hours hours hours hour Additiom 0 16 46 50 N0 addition- 50 0 0 1 2 7 15 26 37 7 EXAMPLE 5 p H Crude root extract of Achyranthes iaponica Nakaii is Slxty silkworms are tested for each expenment' The added to mulberry leaves, and thus obtained feedstufi is fed to silkworms (race: N124 X C124) on the eighth day of the final instar. The results are shown in Table 6.

number of mounting larvae is observed 12 hours after the start of feeding.

TABLE 6 Amount of Mounting number after- Spinning number afteraddition, Number g. of Of silk- 18 23 28 36 48 60 18 23 28 36 42 48 feedstufi worms hours hours hours hours hours hours hours hours hours hours hours hours The results are shown in Table 7.

It is evident that the beneficial effect on the acceleration of mounting is not observed in the case of materials 1 to 6 added.

EXAMPLE 7 0, 125, 250, 500, 1000, 1500, 2000, 3000 and 5000 of 2fl,3fi,14u,20,22 pentahydroxycholest-7-ene-6-one are respectively added to the basal feedstufi in Example 6.

Thus obtained feedstutf is given to silkworms (race: Gunko X Banri) from the 4th day of the final instar.

Two hundred of silkworms are tested for each experiment.

The time of mounting and the quality and quantity of cocoons are observed.

The results are shown in Table 8.

TABLE 8 Amount of Ratio of mounting larvae, Ratio of addition percent, innormal 7/100 g. cocoon, Test, feedstufi 12 hours 18 hours 24 hours percent EXAMPLE 8 2p, 3 S,l4u,20,2Z-pentahydroxycholest-7-en-6-one (compound A) and 218,35,l4a,20,22,25-hexahydroxy cholest-7- en-6-one (compound B) are respectively added to the basal feedstuff of Example 6. Thus obtained feedstufii is given to silkworms (race: Gunko x Banri) of the 3rd day and 4th day of the final instar, respectively. The number of mounting larvae is observed 12 hours and 20 hours after the start of feeding. The results are shown in Tables 9 and 10.

TABLE 9 [Added at the 3rd day of the final instar] Amount of Mounting number addition, Number of after- /1 g. silkworrns Test compounds ieedst tested 12 hours 20 hours No addition 0 60 0 0 A 60 0 0 20 60 0 0 1 About 20 to 30 percent of the silkwonns were dead afterwards.

TABLE [Added at the 4th day of the final instarl Amount of Mounting number addition, Number of after-- 'Y/l g. silkwonns Test compounds teedstufi tested 12 hours 20 hours No addition 0 60 0 0 A 5 60 33 55 20 60 47 60 8 EXAMPLE 9 The experiments inpart I are carried out by adopting as basal stuffs various artificial feedstuifs shown in Part II.

Part I.Effect of claimed compounds on simultaneous mounting of silkworms Experiment 1 2y of2p,3p,l4a, 20, 22,26-hexahydroxycholest-7-en-6- one and 31 of 25,35,14a,20,22,25-hexahydroxycholest-7- en-6-one (totally 57) are added to 1 g. of artificial feedstuff A.

Thus obtained feedstufi is fed on silkworms, Bom byx mori L, (race: Gunko X Band) on seventh day of the final instar.

The silkworms are observed to begin to mount in 17 hours, after feeding, and the time course is shown in Table 11 in comparison with the case of no addition.

en-6-one (totally 107) are added to l g. of artificial feedstuff B.

Thus obtained feedstufi is fed on silkworms, Bombyx mori L., (race; Gunko x Banri) on seventh day of the final instar.

The silkworms are observed to begin to mount in 17 hours, after feeding and the time course is shown in Table 12, in comparison with the case of no addition.

TABLE 12 Number of mounting worms after eding for- Numberof Silkworms 12 17 22 27 32 37 42 Test tested hrs. hrs. hrs. hrs. hrs. hrs. hrs.

Additi0n.-.-.. 300 0 i 52 181 258 282 297 300 N0 addition (control) 300 0 11 72 114 171 204 237 EXPERIMENT 3 2.57 of 25,33,14a,20,22,2S-hexahydroxycholest-7-en-6- one and 5' of 2 9,35,140;,20,22-pentahydroxycholest-7-en- 6-one (totally 7.5'y) are added to 1 g. of artificial feedstufi C.

,Thus obtained feedstufi is fed on silkworms (race: Gunko x Hoshun) on 6th day of the final instar.

The silkworms are observed to begin to mount in 17 hours after feeding and the time course is shown in Table 13, in comparison with the case of no addition.

TABLE 13 Number of mounting worms after feeding for- Numberoi silkworms 12 17 22 27 32 37 42 Test tested hrs. hrs. hrs. hrs. hrs. hrs. hrs.

Addition 300 0 28 91 235 264 287 299 N0 addition (control) 300 0 0 0 0 6 9 52 EXPERIMENT 4 2.5 of 2 8,3 3,l4a,20,22,25-hexahydroxycholest-7-en-6- one, and 7.5 of 2 8,35,140;,20,ZZ-pentahydroxycholest-7- en-6-one (totally 10 are added to 1 g. of artificial feedstufif D.

Thus obtained feedstufl' is fed on silkworms (race; Gunko x Banri) on 6th day of the final instar.

The result is shown in Table 14, in comparison with the case of no addition. e a 7 TABLE 14 TABLE 18 Number of mounting worms after Amount of Number of mounting worms e ing oraddition Number of after feeding for- Numberoi /1 g. silkworms silkworms 12 22 27 32 37 feedstufiH tested 11 hrs. 15 hrs. 19 hrs. 23 hrs. Test tested hrs. hrs. hrs. hrs. hrs. hrs. 5

Control 100 0 3 21 78 AddltlOH 300 0 30 138 266 285 293 9 100 91 100 No addition 1 100 61 90 98 100 (control) 300 0 0 0 0 3 1 99 84 99 20 100 100 10 52.5. eahchfoifjfhe; compounds.d

792.0 0 8 \VO compoun S. EXPERI NT 5 8 107 each of the two compounds.

57 of 218,3 8,l4a,20,22,25-hexahydroxycholest-7-en-6- one is added to 1 g. of artificial feedstufi E. TABLE 19 Thus obtained feedstuff is fed on silkworms (race;

Gunko x Banri) on 7th day of the final instar. 2 3332 Number of Numbfir 0I1l11;t111r?d Worms The result is shown in Table 15, in comparison with the m Silkworm a er ee 1 g case of no addition, ieedstufi H tested 11 hrs. 15 hr 21 hrs. 78 hrs.

TABLE 15 Control.. 0 100 0 5 22 67 E53 1% i33 35. "100 f 92 99 Numb i iii tiii (a) no 100 95 100 Number of 3 20 100 100 silkworms 12 17 22 27 32 97 42 Test tested hrs. hrs. hrs. hrs. hrs. hrs. hrs. 2.57 eac of t tWO p s- 2 57/ each of the two compounds. Addition 300 21 54 63 96 155 265 287 10799911 of t two compounds- No addition (control)- 300 26 30 38 49 66 93 140 EXPERIMENT 9 The compounds of the present invention are, solely or EXPERIMENT 6 in combination, added to artificial feedstuif 1. Thus ob- 9 of 2 3,35,14a,20,22-pentahydroxycholest-7-en-6-one tained feedstuifs are fed on silkworms (race; C131 x tri acetate (M.P. 119-120 C.) is added to 1 g. of artificial N131) 011 7th day of the final instar. The time course of feedstufi F, mounting, as shown in Table 20, demonstrates a compari- Thus obtained feedstuif is fed on silkworms (race; son between the case wherein the compounds of the pres- Gunko x Banri) on 7th day of the final instar. ent invention are added as opposed to the case wherein The result is shown in Table 16, in comparison with the such compounds are not added. case of no addition.

TABLE 16 TABLE 20 Number of mounting worms after Number of matured feeding [or wormsafterfeedmgior- Number of Number of silkworms 12 17 22 27 32 37 42 silkworms 9 12 15 18 Test tested hrs. hrs. hrs. hrs. hrs. hrs. hrs. Test tested hrs. hrs. hrs. hrs,

Addition 300 11 66 151 243 276 299 300 Control (no addition). 100 21 22 33 62 No addition Addition 100 25 36 86 98 (control) 300 23 27 36 57 so 119 Do 100 25 46 90 99 Do. 100 25 57 100 45 no. 100 26 63 100 I 0f 57 of 2B,3B,14a,20 22-penta.hydroxyeho1est-7-en-6-one EXPERIMENT 7 peg f h l l if d 35 14 20 22 t h d h l t 7 7 one o a, -pen a y roxyc 0 es -en-' '7 0f 213,33,149920,2212G'hBXahYdIOXYChQIBSt'7'en'5' one and 2939,1467,20,25- entdn drox onolest-7-en-6-one (toone and 77 of 219,313,14u,20,22,25-hexahydroxycholest-7- tal lr 1 p 9' gg i ggt h d 0 h 1 t 7 6 en-6-one (totally 12 are added to 1 g. of artificial feed- 50 one; 2535;,2025 penmhydroxgcholegt 7 egmDe and 2B,

Thus obtalned feedstufi is fed on silkworms (race, 4% 257 each of 2B,3H,14a;0.2z pentahydmxycholest 7 em Gunko x Banri) on 7th day of the final mstar. The result 3 011 222(1234ah14a,i0g2penfiahytir gxycgolest-7-el6glia; -exayroxyc oes--en- -011e; IS 2 9 m Table 17 m companson with the cas of no pentahydroxycholest-7-en-6-one; 2B,3B,14a.,'20,22,25 hexahyadd1t1on. 55 droxycholest-7-en-6-one; 2B,3B,14a,20,22,2 6-hexahydroxycholest-7-en-6 one and 26,35,1411,20,2 2,252G-heptahydroxycholest- TABLE 17 7-en-6-one (totally 17.5 per 1 g. of feedstufi J.

Number off mounting worms after Numberof EXPERIMENT 10 silkw 12 17 22 27 a2 37 42 Test t s i c l 111's. 1115. hrs. hrs. hrs. hrs. hrs. Various amounts of 2a, Y- Addition 298 23 69 153 242 278 287 296 droxycholest-7-en 6-one are added to feedstuif I. Thus ob- No ddi ion tamed feedstutf 1s fed on sllkworms (race; Gunko x 21 28 35 44 101 135 Banri) on 6th day of the final instar.

The time course of mounting is shown in Table 21. EXPERIMENT s 2B,3 3,14a,20,22 pentahydroxycholest-7-en-6-one (1) TABLE 21 and mixtures of 2,8,319,140L,20,22,26-116XahYdlOXYCh0lCSt-7- Number of mounting worms en-6-one and 2fi,3}3,14oz,20,22,25 hexahydroxycholest-7- a??? of N bar of after feeding en-6-one 2 (3 and 4 are added to artificial feedstufl a ,72: fi 28 32 36 H and 1, respectively. feedstufl K. tested hrs. hrs. hrs. hrs. hl'S.

Thus obtained feedstufi are fed on silkworms (race; 0 300 3 5 18 32 14 C131 x N131) on 7th day of the final instar. g i; 332 33% The time course of mounting is shown in Table 18 and 12 300 122 259 275 294 300 Table 19, respectively.

Water, 1.6 times as much by volume as other components.

Part IL-Some examples of artificial silkworm feedstuff Artificial feed tufi F;

Artificial feedstuff. A: Parts by weight 23332 stgg Mulberry leaf powder 20.0

Defatted soy bean meal 35.0 Defatted Soy bean meal Cellulose powder 15.0 Cellulose 'g i 'g Potato starch 15.0 gotato g ei l 233351 05 @5533: C Campesterol 0.2 Inositol 0.5 Agar Sucrose 10.0 Dlpotasslum hydrogen phosphate 0? Vitamin C 1 5 Choline chloride 0.0 Dihydro streptomycin 0.1 g9 phosphate Water, 1.6 times as much by volume as other a 9 a e "f components. Cltrlc acid 0.1

Water, 1.8 times as much by volume as other Artificial feedstufi B: components.

Mulberry leaf Powder A tifi i 1f d mfiG:

bean meal Mulberry leaf powder 50.0 Cellulose powder 15-0 0 Soy bean meal 150 com starch Cellulose powder 15.0 i Potato starch 10.0 i fi-sitosterol 0.5 Dlpotasslum hydrogen phosphate 0.5 Agar 4 0 Dihydro streptomycin 0-1 Sum; Water, 1.8 times as much by volume as other Vitamin I components Dipotassium hydrogen phosphate 0.15

Artificial feedstuff c; Q P syrbate Mulberry leaf powder 16.0 clmc acid Defatted Soy bean meal 59 Water, 2.0 times as much by volume as other Cellulose powder 8.0 components- Potato Starch Artificial feedstufi H: fi-sitosterol Mulberry leaf powder 50.0 Wessons mixture T a B evve s yeast Sucrose Soy bean meal 10.0 Vitamin mixture Trace c Search 100 Vitamin C Stigmasterol 0.2 IHOSitOl Carboxymethyl cellulose 4.0 Dipotassium hydrogen phosphate 0.5 40 Sucrose 3.0 Choline chloride 005 Vitamin C 1,0 Potassium sorbate Dipotassium hydrogen phosphate 0.5 Citric acid Sodium sorbate 0.15 Water, 2.0 times as much by volume as other Citric acid 15 components- Water, 2.0 times as much by volume as other fi f d components. Arti ci-al ee stu D:

Artlficlal feedstulf I: g leaglpowder n Mulberry leaf powder 20.0

m d Alfalfa meal 5.0 Ce ulose POW er Defatted soy bean meal 40.0 Corn starch 15.0 Rice flour 5D fi'sitosteml Cellulose powder 10.0 i i Corn starch 10.0 f p Oleic acid 1.5 Dlpotassmm hydrogen phosphate 0.5 soy bean sterol 05 Dlhydm stmiptomycm' "T" Carboxymethyl cellulose 4.0 Water, 1.8 tunes as much by volume as other Vitamin mixture Trace components Sucrose 3 0 Artificial feedstutt' E: Mulberry leafeowder 209 Potassium ease-331:3:31133133313111: 0.1 Soy bean casem Water, 1.8 times as much by volume as other Cellulose 40.0 components fl-sitosterol 0.5

Agar 4,0 5 Artificial feedstutf 1:

Vi i i t Trace Defatted soy bean meal 50.0 Sucrose 10,0 Wheat meal 5.0 vit i (3 1.0 Potato starch 10.0 m it l 0,6 Cellulose powder l5 0 Dipotassium hydrogen phosphate 0.4 S Y b en oil 3.0 Choline chloride 0.05

Chloramphenicol 6,1 What is claimed is:

Leucomycin 0.1 1. A method for inducing simultaneous mounting of silkworms, which comprises orally feeding silkworms not earlier than the fourth day of the final instar an eifective 13 amount of a member selected from the group consisting of a compound of the formula wherein the two hydroxyls at positions 2 and 3 are aor fl-oriented, and R is 1,5-dimethylhexyl having 2 to 4 hydroxyl groups, and mixtures of the said compounds in an amount of from about 5 to about 20 gammas per gram of an artificial silkworm feedstuff fed to said silkworms, said amount being determined on a dry basis.

2. A method according to claim 1, wherein the said compound is 2B,3[:?,14cz, 20,22-pentahydroxycholest-7-en- 6-one.

3. A method according to claim 1, wherein the said compound is 2u,3a,14a,20,22-pentahydroxycholest-7-en- 6-one.

4. A method acording to claim 1, wherein the said compound is 213,313,14a,20,22,24-hexahydroxycholest-7-en- 6-one.

5. A method according to claim 1, wherein the said compound is 213,3,8,14a,22,25-pentahydroxycholest-7-en-6- one.

6. A method according to claim 1, wherein the said compound is 2,8,35,l4a,20,22,25-hexahydroxycholest-7-en- 6-one.

7. A method according to claim 1, wherein the said compound is 2 3,3 8,1411,20,22,26-hexahydroxycholest-7-en- 6-one.

8. A method according to claim 1, wherein the said compound is 25,35,14a,20,22,25,26-heptahydroxycholest- 7-en-6-one.

9. A silkworm feeding composition which consists essentially of an artificial silkworm feedstufi and which contains an effective amount of a member selected from the group consisting of a compound of the formula wherein the two hydroxyls at positions 2 and 3 are aor fl-oriented, and R is 1,5-dimethylhexyl having 2 to 4 hydroxyl groups, and mixtures of said compounds, in an amount of from about 5 to about 20 gammas per gram of the silkworm feedstuff, on the dry basis.

10. A silkworm feeding composition according to claim 9, wherein the said compound is 2fi,3fi,14ot,2(),22pentahydroxycholest-7-en-6-one.

11. A silkworm feeding composition according to claim 9, wherein the said compound is 2fl,3fi,l4x,20,22-pentahydroxycholest-7-en-6-one.

12. A silkworm feeding composition according to claim 9, wherein the said compound is 25,3fi,14a,20,22,24hexahydroxycholest-7-en-6-one.

13. A silkworm feeding composition according to claim 9, wherein the said compound is 2/8,3fi,l4a,22,25-pentahydroxycholest-7-en-6-one.

14. A silkworm feeding composition according to claim 9, wherein the said compound is 2fi,3/3,14oz,20,22,25-116X21- hydroxycholest-7-en-6-one.

15. A silkworm feeding composition according to claim 9, wherein the said compound is 25,313,14a,20',22,26-hexahydroxycholest-7-en-6-one.

16. A silkworm feeding composition according to claim 9, wherein the said compound is 2p3,3/3,14u,20,22,25,26- heptahydroxycholest-7-en-6-one.

References Cited UNITED STATES PATENTS 5/1969 Wiechert et a1 260397.2 1/ 1967 Kato et a1. 992

US. Cl. X.R. 424242 Patent No. 3,709,990 Dated January 9, 1973 I t Masaki Kamada and Tetsuo Okauchi It is certified that error appears in the. above-identified patent and that said Letters Patent are hereby corrected as shown below:

Page one, first column, below the line reading "Ser. Nor 38,897"; insert the following:

- Claims priority, Japan application's nos. 79939/66,

December 6, 1966, 81174/66, December 10, 1966, 868/67, December 28, 1966 and 41869/67, June 28, 1967 I Signed sealed this 5th day of March 197E.

( SEAL) Attest:

EDWARD M. FLETCHER, JR. C. MARSHALL DANN Atte sting Officer' Commissioner f Patents 2M PO-105O (10-69) USCOMM-DC 90376-P69 U.S. GOVERNMENT PRINTING OFFICE l9! 0-356-334, 

